皮质酮抑制LPS诱导的小鼠巨噬细胞NLRP3表达及与XO的关系  

Corticosterone inhibits LPS-induced expression of NLRP3 in mouse macrophages and its relation with XO

作　　者：吴玲 田泽丹[1] 陈楠[1] 胡薇 周成富 杜权[1] WU Ling;TIAN Ze-dan;CHEN Nan;HU Wei;ZHOU Cheng-fu;DU Quan(Department of Anesthesiology,The Second Affiliated Hospital,Chongqing Medical University,Chongqing 400010,China)

机构地区：[1]重庆医科大学附属第二医院麻醉科,重庆400010

出　　处：《中国病理生理杂志》2018年第4期693-698,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81172122);重庆市医学科研计划项目(No.20121130)

摘　　要：目的:探讨皮质酮(CORT)对脂多糖(LPS)诱导的小鼠巨噬细胞核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)表达的抑制作用及与黄嘌呤氧化酶(XO)的关系。方法:用LPS构建小鼠巨噬细胞RAW 264.7的炎症模型。运用不同浓度(0~900μg/L)的CORT处理巨噬细胞后提取细胞总蛋白,Western blot法检测细胞NLRP3和caspase-1的蛋白水平;按处理因素分组为:对照组、LPS组、LPS+CORT组和LPS+别嘌呤醇(allopurinol)组,分别于0、0.5、1、1.5和2 h提取细胞成分,运用real-time PCR和Western blot法检测细胞NLRP3和XO的m RNA和蛋白水平。结果:高于700μg/L的CORT可显著抑制LPS诱导的巨噬细胞中NLRP3的表达及caspase-1的活化(P<0.05);与LPS组相比,LPS+CORT在下调LPS诱导的巨噬细胞NLRP3表达的同时抑制XO的表达(P<0.05),LPS+allopurinol组巨噬细胞NLRP3的表达减少(P<0.05)。结论:较高浓度的CORT能抑制LPS诱导的小鼠巨噬细胞NLRP3的表达,而CORT的抑制作用可能与其下调XO的表达水平有关。AIM:To investigate the inhibitory effect of corticosterone(CORT)on lipopolysaccharide(LPS)-induced expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)and its relation with xanthine oxidase(XO).METHODS:An inflammatory model of mouse macrophage RAW 264.7 was established by stimulating with LPS.Total cellular protein was extracted after the macrophages were treated with CORT at different concentrations(0~900μg/L).The protein levels of NLRP3 and caspase-1 were determined by Western blot.According to the treatments,the macrophages were divided into control group,LPS group,LPS+CORT group and LPS+allopurinol group.Cell components were extracted at 0,0.5,1,1.5 and 2 h.The protein levels of NLRP3 and XO were determined by Western blot,and the mRNA expression of NLRP3 and XO was detected by real-time PCR.RESULTS:CORT at 700μg/L and above significantly inhibited the expression of NLRP3 and the activation of caspase-1 in the macrophages induced by LPS(P<0.05).Compared with LPS group,the expression of NLRP3 and XO in LPS+CORT group was inhibited(P<0.05),and the expression of NLRP3 in LPS+allopurinol group was also reduced(P<0.05).CONCLUSION:High concentration of CORT inhibits the expression of NLRP3 in LPS-induced mouse macrophages,which is associated with XO.The inhibitory effect of CORT may be related to the reduction of XO expression.

关 键 词：脂多糖 皮质酮 核苷酸结合寡聚化结构域样受体蛋白3 黄嘌呤氧化酶 

分 类 号：R364.5[医药卫生—病理学] R363.2[医药卫生—基础医学]